Biochem Pharmacol, 1998 May 1, 55(9), 1339 - 46 Ras, superoxide and signal transduction; Irani K et al.; The superoxide anion has been associated with the bactericidal activity of phagocytes . Produced by an enzymatic complex, NADPH oxidase, bactericidal superoxide is released within phagolysosomes where bacteria are being degraded . The activity of NADPH oxidase is regulated by Rac, a small GTP binding protein of the Ras family . Recent evidence indicates that, in addition to its bactericidal activity, superoxide seems to function as a signal-transduction messenger, mediating the downstream effects of Ras and Rac in nonphagocytic cells . As such, superoxide contributes to the unchecked proliferation of Ras-transformed cells . In the nitric oxide (NO) system, low concentrations of NO transduce signals within vessels and neurons, while high concentrations of NO can produce damage to cells and microorganisms . By analogy, superoxide and probably other oxidants serve as messengers at low concentrations, while larger amounts are required for inducing damage . The activity of oxidants as messengers opens new avenues for pharmacological intervention against Ras-mediated pathways in mammalian cells. Scand J Immunol, 1999 Feb, 49(2), 193 - 6 Mannan-binding lectin deficiency is associated with unexplained recurrent miscarriage; Christiansen OB et al.; Mannan-binding lectin (MBL) is a plasma protein which, upon binding to microbial carbohydrate structures, elicits activation of the complement system . The level of MBL is genetically determined . It has been reported that the frequency of low plasma levels of MBL is increased in patients with unexplained recurrent miscarriages (RM) . In the present study plasma MBL levels were determined in 146 Danish women with RM and 41 of their husbands together with 49 Scottish RM women and 41 of their husbands . In both countries MBL levels were also investigated in a total of 444 controls . Based on the control data, a cut-off MBL level < 50 ng/ml was selected to define MBL deficiency . The typical odds ratio for MBL deficiency among female patients in the two populations was 1.68 (95% confidence limits 1.01-2.80, P<0.05) whereas it was 1.57 (95% confidence limits 0.72-3.42, not significant) for the male partners of the patients . There was a significant correlation between the frequency of MBL deficiency in RM women and the number of previous miscarriages (P < 0.01), whereas no such correlation was found in the husbands . The results indicate that maternal MBL deficiency is associated with RM . Maternal MBL deficiency might impair the immune defence against microorganisms at the feto-maternal interface. Ann Intern Med, 1999 Jan 19, 130(2), 126 - 30 Compliance with handwashing in a teaching hospital . Infection Control Program; Pittet D et al.; BACKGROUND: Transmission of microorganisms from the hands of health care workers is the main source of cross-infection in hospitals and can be prevented by handwashing . OBJECTIVE: To identify predictors of noncompliance with handwashing during routine patient care . DESIGN: Observational study . SETTING: Teaching hospital in Geneva, Switzerland . PARTICIPANTS: Nurses (66%), physicians (10%), nursing assistants (13%), and other health care workers (11%) . MEASUREMENTS: Compliance with handwashing . RESULTS: In 2834 observed opportunities for handwashing, average compliance was 48% . In multivariate analysis, noncompliance was higher among physicians (odds ratio {OR}, 2.8 {95% CI, 1.9 to 4.1}), nursing assistants (OR, 1.3 {CI, 1.0 to 1.6}), and other health care workers (OR, 2.1 {CI, 1.4 to 3.2}) than among nurses and was lowest on weekends (OR, 0.6 {CI, 0.4 to 0.8}) . Noncompliance was higher in intensive care than in internal medicine units (OR, 2.0 {CI, 1.3 to 3.1}), during procedures that carry a high risk for contamination (OR, 1.8 {CI, 1.4 to 2.4}), and when intensity of patient care was high (compared with < or = 20 opportunities for handwashing per hour of care, 21 to 40 opportunities: OR, 1.3 {CI, 1.0 to 1.7}; 41 to 60 opportunities: OR, 2.1 {CI, 1.5 to 2.9}; and > 60 opportunities: OR, 2.1 {CI, 1.3 to 3.5}) . CONCLUSIONS: Compliance with handwashing was moderate . Variation across hospital ward and type of health care worker suggests that targeted educational programs may be useful . Even though observational data cannot prove causality, the association between noncompliance and intensity of care suggests that understaffing may decrease quality of patient care. J Chromatogr B Biomed Sci Appl, 1999 Feb 5, 722(1-2), 263 - 78 Methods for the differentiation of microorganisms; Busch U et al.; Advances in analytical and diagnostic assays based on novel nucleic acid analyses techniques have revolutionized the application of molecular differentiation of microorganisms . Phenotypic typing schemes are now broadly supplemented by new genotyping methods which allow a more refined and detailed differentiation of closely related microorganisms, bacterial strains, isolates and pathogens on the DNA level . Bio-, sero- and phagetyping, antibiotic susceptibility tests, immunoblotting as well as multilocus enzyme- or polyacrylamide gel electrophoresis are now supported by the analysis of plasmid or chromosomal DNA restriction profiles, ribotyping, pulsed-field gel electrophoresis and polymerase- or ligase-chain reaction-based methods or direct sequencing technique to differentiate microorganisms . Some of these molecular techniques are also used in the field of virology to analyse and differentiate closely related sub- or genotypes . Few examples for the analysis and investigation of these usually small genomes will also be given. Curr Opin Microbiol, 1998 Dec, 1(6), 643 - 8 Two-component signal transduction systems in eukaryotic microorganisms; Loomis WF et al.; Conserved signal transduction pathways that use phosphorelay from histidine kinases through an intermediate transfer protein (H2) to response regulators have been found in a variety of eukaryotic microorganisms . Several of these pathways are linked to mitogen-activated protein kinase cascades . These networks control different physiological responses including osmoregulation, cAMP levels and cellular morphogenesis. Curr Opin Microbiol, 1998 Jun, 1(3), 278 - 85 The value of microbial diversity; Hunter-Cevera JC; In the past few years, due to the use of molecular methods, our knowledge of microbial diversity has increased dramatically, not only from a phylogenetic and taxonomic perspective but also from an ecological basis . We now know that microorganisms exist in every conceivable place on Earth, even in extreme environments . Temperature may be the only limitation as to where they can and cannot exist and/or function . As more small subunit rDNA sequence information becomes available there is a real need to start turning the information into knowledge that can be applied to better elucidate and understand structure-function relationships within ecosystems, develop new culturing methods, and discover new products and processes . It has been stated on numerous occasions that the 21(st) century is the century for biology . Within that context, we must address the real value of microbial diversity. Curr Opin Microbiol, 1998 Jun, 1(3), 319 - 29 Combinatorial biosynthesis for new drug discovery; Hutchinson CR; Combinatorial biosynthesis involves interchanging secondary metabolism genes between antibiotic-producing microorganisms to create unnatural gene combinations or hybrid genes if only part of a gene is exchanged . Novel metabolites can be made by both approaches, due to the effect of a new enzyme on a metabolic pathway or to the formation of proteins with new enzymatic properties . The method has been particularly successful with polyketide synthase (PKS) genes: derivatives of medically important macrolide antibiotics and unusual polycyclic aromatic compounds have been produced by novel combinations of the type I and type II PKS genes, respectively . Recent extensions of the approach to include deoxysugar biosynthesis genes have expanded the possibilities for making new microbial metabolites and discovering valuable drugs through the genetic engineering of bacteria. Curr Opin Microbiol, 1998 Jun, 1(3), 291 - 5 Barophiles: deep-sea microorganisms adapted to an extreme environment; Horikoshi K; The deep-sea environment is characterized by high pressure and low temperature but in the vicinity of hydrothermal vents regions of extremely high temperature exist . Deep-sea microorganisms have specially adapted features that enable them to live and grow in this extreme environment . Recent research on the physiology and molecular biology of deep-sea barophilic bacteria has identified pressure-regulated operons and shown that microbial growth is influenced by the relationship between temperature and pressure in the deep-sea environment. Curr Opin Microbiol, 1998 Jun, 1(3), 352 - 8 In situ methods for assessment of microorganisms and their activities; Amann R et al.; Recent technical developments in the field of molecular biology and microsensors are beginning to enable microbiologists to study the abundance, localization and activity of microorganisms in situ . The various new methods on their own bear high potential but it is the combination of studies on structure and function of microbial communities that will yield the most detailed insights in the way microorganisms operate in nature. Biochem Biophys Res Commun, 1999 Mar 5, 256(1), 231 - 4 Metabolic properties of normal and mutant mannan-binding proteins in mouse plasma; Naito H et al.; Human mannan-binding protein (MBP) is a serum lectin involved in innate immunity . MBP activates the complement pathway through its interaction with mannose-rich carbohydrates on various microorganisms and a common opsonic defect has been shown to be associated with a low serum concentration of MBP . This low serum concentration is closely associated with a single base mutation in codon 52, 54 or 57 of the human MBP gene, which results in a change of Arg52 to Cys, Gly54 to Asp, or Gly57 to Gln, respectively, in the collagen-like region of the molecule and prevents the formation of higher oligomers . However, the mechanism underlying the low serum concentration in such patients is completely unknown . The levels of protein synthesis and secretion of the normal and mutant MBPs seem to be similar according to our previous in vitro results . In this study, we examined the plasma clearance of the normal and mutant human (Gly54Asp) MBPs in mice, and found that the half-life of the mutant MBP is about half that of the normal MBP, explaining in part the difference in the plasma levels between the two types of MBP . Scand J Infect Dis, 1998, 30(5), 495 - 9 Low levels of IgG antibodies against pneumocystis carinii among HIV-infected patients; Laursen AL et al.; IgG antibodies against Pneumocystis carinii (P . carinii) were detected by an ELISA method using urea-extracted material from human and rat P . carinii as the antigen . Carbohydrate formed a major part of the antigen responsible for reactivity in the ELISA assay, since periodate treatment reduced the reactivity of most sera tested . Cross-reactivity between human and rat P . carinii was detected . However, human serum recognized antigens specific for human P . carinii . With the ELISA method IgG antibody levels were compared between blood donors (n = 40), asymptomatic HIV-antibody positive patients (n = 30) and AIDS patients with (n=22) and without previous P . carinii pneumonia (PCP) (n=21) . HIV-infected patients had significantly lower antibody reactivity against the microorganism compared with blood donors . Among HIV-antibody positive patients the highest antibody reactivity was seen in PCP patients . The antibody response to PCP was impaired, since an equal number of patients had an increase and a decrease in antibody reactivity . In conclusion, carbohydrate formed an important part of the P . carinii immunogenic antigen . Cross-reactivity between rat and human P . carinii was demonstrated, but reactivity was somewhat lower using antigen from rats . The antibody level was lower in HIV-infected patients and the ability to mount an antibody response to the infection was impaired, suggesting that the poor antibody response may contribute to the liability of HIV-infected patients to have PCP. Scand J Infect Dis, 1998, 30(5), 481 - 3 A blood micro-culture system for the diagnosis of bacteremia in pediatric patients; Solorzano-Santos F et al.; The aim of this study was to evaluate the utility of a volume-modified blood culture system to diagnose bacteremia in newborns and infants . A total of 793 paired blood cultures, obtained from 464 patients (173 newborns and 291 infants), were analyzed . Vacutainer tubes containing 18 ml supplemented peptone broth sodium-polyanethol-sulfonate were used as the gold standard, in comparison with a blood micro-culture system containing 1.8 ml of the broth . Prior to antibiotic treatment, 2.2 ml of blood was obtained from each patient; 0.2 ml was inoculated in a blood micro-culture tube and 2 ml in a routine tube . Sensitivity, specificity and predictive values were calculated . Microorganisms were isolated in 153 standard blood culture tubes and 151 blood micro-culture tubes . The sensitivity of the blood micro-culture system was 95%, specificity 99% and positive and negative predictive values 96% and 99% respectively . The sensitivity and specificity of blood micro-culture in neonates and infants is high . We recommend that this system be used for the diagnosis of bacteremia in newborns and infants in laboratories where manual systems are still in use. Biosci Biotechnol Biochem, 1999 Jan, 63(1), 1 - 9 Microbial desulfurization of organic sulfur compounds in petroleum; Ohshiro T et al.; Sulfur removal from petroleum is important from the standpoint of the global environment because the combustion of sulfur compounds leads to the production of sulfur oxides, which are the source of acid rain . As the regulations for sulfur in fuels become more stringent, the existing chemical desulfurizations are coming inadequate for the "deeper desulfurization" to produce lower-sulfur fuels without new and innovative processes . Biodesulfurization is rising as one of the candidates . Several microorganisms were found to desulfurize dibenzothiophene (DBT), a representative of the organic sulfur compounds in petroleum, forming a sulfur-free compound, 2-hydroxybiphenyl . They are promising as biocatalysts in the microbial desulfurization of petroleum because without assimilation of the carbon content, they remove only sulfur from the heterocyclic compounds which is refractory to conventional chemical desulfurization . Both enzymological and molecular genetic studies are now in progress for the purpose of obtaining improved desulfurization activity of organisms . The genes involved in the sulfur-specific DBT desulfurization were identified and the corresponding enzymes have been investigated . From the practical point of view, it has been proved that the microbial desulfurization proceeds in the presence of high concentrations of hydrocarbons, and more complicated DBT analogs are also desulfurized by the microorganisms . This review outlines the progress in the studies of the microbial desulfurization from the basic and practical point of view. Khirurgiia (Mosk), 1999, (1), 12 - 3 {Complications of sorption-application therapy of purulent wounds}; Pavlov VV et al.; A retrospective analysis of 187 case records was made for elucidation of the frequency of development of complications in sorption-applicational treatment of festering wounds . It was revealed that late after strewing sterile sorbent into the wound in 6.4% of patients the development of fistulas and going out of preparation granules though fistula occurred . It is concluded, that sorbent's granules incapsulate together with living microorganisms adsorbed on their surface . For prevention of this complication the corbent is recommended to be introduced into the wound in small porous containers, which provide quick replacement of the preparation during wound dressing and its complete elimination in the end of healing. PDA J Pharm Sci Technol, 1998 Nov-Dec, 52(6), 344 - 5 Neutralization of beta-lactam antibiotics in an environmental monitoring medium; Winely C et al.; The neutralizing ability of Difco Bacto Penase, BBL Penicillinase, and Genzyme beta-lactamase for penicillin G, oxacillin, cephalothin, cefazolin, ceftazidime, ceftriaxone, cefotaxime, imipenem, and meropenem was determined in an agar medium . Trypticase soy agar plates (20 mL) containing 200 MU/L Difco Penase, 200 MU/L BBL Penicillinase, or 2 vials/L of Genzyme beta-lactamase were dosed with 20, 50, and 100 micrograms of each antibiotic and then inoculated (50-100 CFU/plate) with a susceptible microorganism . Percentage recoveries of the organism were calculated after incubation . Genzyme beta-lactamase effectively neutralized 100 micrograms of all the antibiotics except ceftazidime (32% recovery with 20 micrograms) . The two penicillinases effectively neutralized 100 micrograms of penicillin G, oxacillin, cephalothin, and cefazolin; were slightly effective against ceftriaxone and cefotaxime (20-57% recovery with 20 micrograms); and totally ineffective against ceftazidime, imipenem, and meropenem . This investigation resulted in a useful procedure for qualifying and/or selecting a beta-lactamase for use in environmental monitoring medium to neutralize a particular beta-lactam. Appl Environ Microbiol, 1999 Mar, 65(3), 893 - 7 Relationship between glycosyl hydrolase inventory and growth physiology of the hyperthermophile Pyrococcus furiosus on carbohydrate-based media; Driskill LE et al.; Utilization of a range of carbohydrates for growth by the hyperthermophile Pyrococcus furiosus was investigated by examining the spectrum of glycosyl hydrolases produced by this microorganism and the thermal labilities of various saccharides . Previously, P . furiosus had been found to grow in batch cultures on several alpha-linked carbohydrates and cellobiose but not on glucose or other beta-linked sugars . Although P . furiosus was not able to grow on any nonglucan carbohydrate or any form of cellulose in this study (growth on oat spelt arabinoxylan was attributed to glucan contamination of this substrate), significant growth at 98 degrees C occurred on beta-1,3- and beta-1,3-beta-1,4-linked glucans . Oligosaccharides generated by digestion with a recombinant laminarinase derived from P . furiosus were the compounds that were most effective in stimulating growth of the microorganism . In several cases, periodic addition of beta-glucan substrates to fed-batch cultures limited adverse thermochemical modifications of the carbohydrates (i.e., Maillard reactions and caramelization) and led to significant increases (as much as two- to threefold) in the cell yields . While glucose had only a marginally positive effect on growth in batch culture, the final cell densities nearly tripled when glucose was added by the fed-batch procedure . Nonenzymatic browning reactions were found to be significant at 98 degrees C for saccharides with degrees of polymerization (DP) ranging from 1 to 6; glucose was the most labile compound on a mass basis and the least labile compound on a molar basis . This suggests that for DP of 2 or greater protection of the nonreducing monosaccharide component may be a factor in substrate availability . For P . furiosus, carbohydrate utilization patterns were found to reflect the distribution of the glycosyl hydrolases which are known to be produced by this microorganism. Antimicrob Agents Chemother, 1999 Mar, 43(3), 714 - 6 Lactic acid polymers as biodegradable carriers of fluoroquinolones: an in vitro study; Kanellakopoulou K et al.; A biodegradable polymer of DL-dilactide that facilitates release of ciprofloxacin or pefloxacin at levels exceeding MICs for the causative microorganisms of chronic osteomyelitis is described . Duration and peak of release were found to depend on the molecular weight of the polymer . Its characteristics make it promising for treating chronic bone infections. Wien Klin Wochenschr, 1998 Dec 23, 110(24), 859 - 62 Genetic studies in Borrelia burgdorferi; Rosa P et al.; Borrelia burgdorferi, the agent of Lyme disease, has recently joined a growing number of micro-organisms for which the entire genomic sequence is known . Despite this wealth of information, little is known about the contribution of specific spirochetal components to the pathogenesis of Lyme disease or their function in the normal life cycle of the organism . This discrepancy is due in part to the lack of a well-developed genetic system in B . burgdorferi, which in turn can be attributed to its relatively recent isolation and the dissimilarity of Borrelia from other genetically tractable bacteria . We are interested in several plasmid-encoded gene products in B . burgdorferi that may play a role in sensing and adaptation to the different environments the spirochete encounters as it completes an infectious cycle between the tick vector and the mammalian host . We are developing genetic tools with which to test the roles of specific B . burgdorferi gene products in the transmission cycle in an animal model of Lyme disease . We have demonstrated targeted gene inactivation by allelic exchange, using the gyrBr gene encoding coumermycin-resistant topoisomerase as a selectable marker . Spirochetes are transformed by electroporation and coumermycin-resistant colonies are screened by PCR for allelic exchange at the targeted locus . We have successfully inactivated several genes of interest in the type strain B31 . We are investigating the utility of additional antibiotic resistance genes as selectable markers in B . burgdorferi . Targeted gene inactivation is a powerful tool with which to investigate the role of particular proteins in the basic biology and virulence of a pathogenic microorganism . We have made significant advances in our ability to genetically manipulate B . burgdorferi in order to address these issues . However, the available methods are incomplete and far from routine . We are currently improving existing methods as well as developing additional genetic tools with which to augment genetic studies in B . burgdorferi. Wien Klin Wochenschr, 1998 Dec 23, 110(24), 856 - 8 Natural history of Borrelia burgdorferi sensu lato; Gern L et al.; Lyme borreliosis is a zoonosis: its causative agent, B: burgdorferi, circulates between ticks and a large range of vertebrates . Identification of the hosts which are responsible for the infection of the vectors is extremely important to determine the potential risk of infection in an habitat . Various small mammals and bird species are considered reservoirs for the Lyme disease spirochetes . Grey and red squirrels, hedgehogs as well as hares and rabbits can develop an infection and transmit B . burgdorferi sensu lato to feeding ticks . In Eurasian endemic areas, many different Borrelia species circulate between ticks and vertebrate hosts . Studies have shown that European and Asian genospecies are associated with specific groups of vertebrate hosts, such as B . valaisiana and B . garinii with birds, B . afzelii with small mammals and B . burgdorferi ss and B . afzelii with red squirrels . However, such associations are not always observed as in Japan where B . garinii, B . afzelii and unidentified Borrelia species are found in small mammals . Some enzootic cycles involving tick species which do not feed at all on humans or which rarely feed on humans have been described in Europe and USA . It is likely that many existing enzootic foci have yet to be discovered . The circulation of B . burgdorferi in silent foci does not have important implications for human health, but it demonstrates the complexity of the ecology of this microorganism and the variety of ecological niches this spirochete can occupy. J Food Prot, 1999 Feb, 62(2), 128 - 32 Inhibition of growth of Escherichia coli O157:H7 in fresh radish (Raphanus sativus L.) sprout production by calcinated calcium; Bari ML et al.; The inhibitory effect of calcinated calcium on the growth of Escherichia coli O157:H7 during fresh radish (Raphanus sativus L.) sprout production was studied . It was revealed that the addition of 0.4% (wt/vol) calcinated calcium into radish sprouting medium which was artificially contaminated with E . coli O157:H7 (3.0 to 3.2 log CFU/ml) completely inhibited the growth or inactivated the microorganism . When radish seed extract was used instead of radish sprout production, the same extent of growth inhibition or inactivation was observed with much lower amounts (0.07%) of calcinated calcium under similar experimental conditions . The findings suggested that calcinated calcium may be useful to control E . coli O157:H7 contamination during the production of radish sprouts. Yeast, 1999 Jan 15, 15(1), 23 - 33 Saccharomyces cerevisiae IRR1 protein is indirectly involved in colony formation; Kurlandzka A et al.; The ability of a microorganism to adhere to a solid support and to initiate a colony is often the first stage of microbial infections . To date, studies on S . cerevisiae cell-cell and cell-solid support interactions concerned only cell agglutination during mating and flocculation . Colony formation has not been studied before probably because this species is not pathogenic . However, S . cerevisiae can be a convenient model to study this process, thanks to well-developed genetics and the full knowledge of its nucleotide sequence . A preliminary characterization of the recently cloned essential IRR1 gene indicated that it may participate in cell-cell/substrate interactions . Here we show that lowering the level of expression of IRR1 (after fusion with a regulatory catalase A gene promoter) affects colony formation and disturbs zygote formation and spore germination . All these processes involve cell-cell or cell-solid support contacts . The IRR1 protein is localized in the cytosol as verified by immunofluorescence microscopy, and confirmed by cell fractionation and Western blotting . This indicates that Irr1p is not directly involved in the cell-solid support adhesion, but may be an element of a communication pathway between the cell and its surroundings. Yeast, 1999 Jan 15, 15(1), 11 - 22 Cloning, sequence analysis and overexpression of a Saccharomyces cerevisiae endopolygalacturonase-encoding gene (PGL1); Gognies S et al.; Only a few yeast strains produce pectin-degrading enzymes such as pectin esterases and depolymerases (hydrolases and lyases) . Strain SCPP is the only known Saccharomyces strain to produce these pectinases . One of these pectolytic enzymes . PGL1-encoded endopolygalacturonase (EC 3.2.1.15), hydrolyses the alpha-1,4-glycosidic bonds within the rhamnogalacturonan chains in pectic substances . This paper presents the cloning and sequencing of the first S . cerevisiae gene involved in pectin degradation . Few differences were found between the two deduced amino acid sequences encoded by PGL1-1 from a pectolytic (PG+) strain (SCPP) and PGL1-2 from a non-pectolytic (PG-) strain (X2180-1B) . Similarities were found with other polygalacturonases from plants and other microorganisms . Of the two S . cerevisiae genes, only the one isolated from strain SCPP was able, by overexpression, to confer endopolygalacturonase activity to a laboratory strain of S . cerevisiae . Overexpression of PGL1-1 gene in a non-pectolytic strain resulted in halo formation on polygalacturonic acid-containing agar plates stained with ruthenium red. Circulation, 1999 Feb 23, 99(7), 879 - 82 Detection of Chlamydia pneumoniae but not cytomegalovirus in occluded saphenous vein coronary artery bypass grafts; Bartels C et al.; BACKGROUND: A causal relation between atherosclerosis and chronic infection with Chlamydia pneumoniae and/or cytomegalovirus (CMV) has been suggested . Whether the unresolved problem of venous coronary artery bypass graft occlusion is related to infection with C pneumoniae and/or CMV has not been addressed . METHODS AND RESUTLS: Thirty-eight occluded coronary artery vein grafts and 20 native saphenous veins were examined . Detection of C pneumoniae DNA was performed by use of nested polymerase chain reaction (PCR) . Homogenisates from the specimen were cultured for identification of viable C pneumoniae . Both conventional PCR and quantitative PCR for detection of CMV DNA were applied . Differential pathological changes (degree of inflammation, smooth muscle cell proliferation {MIB-1}) were determined and correlated to the detection of both microorganisms . C pneumoniae DNA could be detected in 25% of occluded vein grafts . Viable C pneumoniae was recovered from 16% of occluded vein grafts . Except for 1 native saphenous vein, all control vessels were negative for both C pneumoniae detection and culture . All pathological and control specimens were negative for CMV DNA detection . Pathological changes did not correlate with C pneumoniae detection . CONCLUSIONS: Occluded aorto-coronary venous grafts harbor C pneumoniae but not CMV . The detection of C pneumoniae in occluded vein grafts warrants further investigation. J Chromatogr B Biomed Sci Appl, 1999 Jan 8, 721(1), 31 - 7 Surface characterization and on-line activity measurements of microorganisms by capillary zone electrophoresis; Torimura M et al.; Capillary zone electrophoresis (CZE) was applied to the electrophoretic characterization for microorganisms . The electrophoretic peaks detected using light scattering phenomena were characteristic of the microorganisms used . The electrophoretic mobility (mu) evaluated by CZE was in good agreement with that obtained by classical electrophoresis of microorganisms . The migration time was reproducible and depended on the ionic strength (I) . Analysis of the mu vs . I relationship provided information regarding the charge density and the hardness of the microbial cell surface . The redox enzymatic activity of microorganisms was also evaluated by CZE using a running buffer containing a corresponding substrate and an appropriate exogenous electron acceptor . A decrease in the concentration of the electron acceptor due to microbial activity can be simultaneously monitored during the electrophoretic process without significant modification of the CZE instrument . Effects of some chemical treatments of microbial cells were also studied using this technique. J Clin Lab Anal, 1999, 13(1), 1 - 4 Reliability of low-avidity IgG and of IgA in the diagnosis of primary infection by rubella virus with adaptation of a commercial test; Gutierrez J et al.; The detection of IgA and low-avidity IgG and antibodies in serum is a potentially useful marker of recent infection by a microorganism . We studied the reliability of IgG avidity and presence of IgA for the diagnosis of recent acute infection by rubella virus . Low-avidity IgG (Avy-EIA test) was determined with a modified commercial test using 8 molar urea (indirect ELISA, DiaSorin, Italy) and IgA was determined with a homemade indirect ELISA test . Twenty-five patients with recent primary infection by rubella virus (group I) and 50 healthy subjects (group II) were studied . In group I low-avidity IgG varied between 100 and 0% (67.3+/-21.8%); IgA was present in 24 patients (96%) . In group II low-avidity IgG varied from 50.4 to 0% (19.8+/-16.9%) . IgA was present in 2 subjects (4%) . The sensitivity of the Avi-EIA and the IgA test was 92 and 96%, respectively; specificity was 100 and 96%, respectively . We conclude that both low-avidity IgG and IgA tests are helpful and reliable for the diagnosis of recent primary infection. Phys Rev A, 1991 Oct 15, 44(8), 5279 - 91 Evolution of bioconvective patterns in variable gravity; Noever DA; Measurements are reported of the evolution of bioconvective patterns in shallow, dense cultures of microorganisms subjected to varying gravity . Various statistical properties of this random, quasi-two-dimensional structure have been found: Aboav's law is obeyed, the average vertex angles follow predictions for regular polygons, and the area of a pattern varies linearly with its number of sides . As gravity varies between 1 g and 1.8g (g = 9.8 m s-1), these statistical properties continue to hold despite a tripling of the number of polygons and a reduced average polygon dimension by a third . This work compares with experiments on soap foams, Langmuir monolayer foams, metal grains, and simulations. J Immunol Methods, 1999 Jan 1, 222(1-2), 135 - 44 High-level and effective production of human mannan-binding lectin (MBL) in Chinese hamster ovary (CHO) cells; Ohtani K et al.; We have developed a high-expression system of recombinant human mannan-binding lectin (MBL) with CHO cells . Geneticin-resistant transformants harboring human MBL cDNA in the expression vector pNOW/CMV-A were screened by immunoblot analysis for secretion of recombinant MBL . Cloning and selection by both geneticin and methotrexate resulted in the production of recombinant MBL to a final concentration of 128.8 microg/ml in media after four days of culture . SDS-PAGE and gel-filtration analyses showed that recombinant MBL is characterized by two lower-order oligomeric structures (apparent molecular weights: 1150 kDa and 300 kDa) compared to native MBL (apparent molecular weight: 1300 kDa) . The recombinant human MBL has both sugar-binding and complement activation activity and, like native MBL, can inhibit hemagglutination of influenza A virus . Lectin blots with recombinant MBL indicate that it can bind such microorganisms as HIV and influenza virus suggesting that it might inhibit their infection of hosts . This high-level expression of human MBL with the full range of biological activity will be useful for studies on the immunological role of MBL in humans. Toxicol Lett, 1998 Dec 28, 102-103, 283 - 7 Immune and inflammatory responses in the CNS: modulation by astrocytes; Aschner M; Because the skull bones, the cerebrospinal fluid, the blood-brain barrier (BBB), and the meninges effectively shield the central nervous system from other tissues, it was proposed that the brain is an 'immunologically privileged' organ . However, with recent evidence that in response to invasion by microorganisms, resident cells, such as astrocytes and microglia can fully mount an immune response, this long-standing view has been rethought and revised . Over the last two decades, both astrocytes and microglia have been shown to secrete numerous cytokines, and, therefore, it is presently widely accepted that these cells actively participate in an integrative communicative pathway between resident immune cells of the CNS and those of the periphery . While clearly implicated in the initiation, maintenance, and suppression of immune responses, cytokines produced by these cells (e.g . astrocytes and microglia), as well as the responses of these cells to cytokines produced elsewhere, has also been shown to propagate CNS damage . Therefore the potential involvement of these cells in neurodegenerative disorders has been raised and subjected to intense experimentation . The objective of this synopsis is to review the role played by astrocytes in the initiation and modulation of immune responses. J Exp Biol, 1999 Mar, 202 ( Pt 6), 645 - 53 Inducible NO synthase: role in cellular signalling; Beck KF et al.; The discovery of endothelium-derived relaxing factor and its identification as nitric oxide (NO) was one of the most exciting discoveries of biomedical research in the 1980s . Besides its potent vasodilatory effects, NO was found under certain circumstances to be responsible for the killing of microorganisms and tumour cells by activated macrophages and to act as a novel, unconventional type of neurotransmitter . In 1992, Science picked NO as the 'Molecule of the Year', and over the past years NO has become established as a universal intercellular messenger that acutely affects important signalling pathways and, on a more long-term scale, modulates gene expression in target cells . These actions will form the focus of the present review. Diagn Microbiol Infect Dis, 1999 Jan, 33(1), 1 - 6 The detection of positive blood cultures by the AccuMed ESP-384 system: the clinical significance of three-day testing; Han XY et al.; A total of 805 positive blood cultures obtained over a 4-month period during 1997 by the AccuMed ESP-384 system were evaluated and compared with 471 positive blood cultures obtained during 1989 . Of the 805 microorganisms isolated in 5-day culture and testing by the ESP-384, 671 (83.4%) were detected within the first 2 days and 758 (94.2%) within 3 days . There were 47 (5.7%) microorganisms detected in culture Days 4 and 5 from 42 patients and review of the medical records from these patients demonstrated that no significant changes in clinical management were instituted as a result of detection and identification of these isolates . These 47 organisms were considered to be zero to equivocal clinical relevance . These data suggest that a 3-day routine blood culture incubation protocol with the ESP-384 system may be sufficient when considering detection rate, clinical relevance, and cost-effectiveness . The microbial spectrum and relative frequencies in this study were found to be similar to those of positive blood cultures obtained during 1989. Klin Khir, 1998, (6), 23 - 6 {Risk factors determining the outcome of a diffuse peritonitis of appendicular genesis in children}; Moskalenko VZ et al.; The peculiarities of pre-, intra- and early postoperative period course in 108 children, operated on for diffuse and general peritonitis of appendicitis origin, are studied . Most significant 34 prognostic factors for the disease outcome are choosed . The leading factors are the disease course duration, general condition of the patient while hospitalization, the vegetative disorders presence, the intestinal paresis degree, the biochemical inductors of stress contents, the peritoneal exudate character, the kind and composition of microorganisms in it, the character of an early postoperative period course. Mol Microbiol, 1998 Dec, 30(5), 1003 - 15 Identification of a 47 kDa fibronectin-binding protein expressed by Borrelia burgdorferi isolate B31; Probert WS et al.; The attachment of pathogenic microorganisms to host cells and tissues is often mediated through the expression of surface receptors recognizing components of the extracellular matrix (ECM) . Here, we investigate the ability of Borrelia spirochaetes to bind the ECM constituent, fibronectin . Borrelia lysates were separated by SDS-PAGE, transferred to nitrocellulose and probed with alkaline phosphatase-labelled fibronectin (fibronectin-AP) . Five of six Borrelia species and four of eight B . burgdorferi sensu lato isolates expressed one or more fibronectin-binding proteins . Borrelia burgdorferi isolate B31 expressed a 47 kDa (P47) fibronectin-binding protein that was localized to the outer envelope based on susceptibility to proteinase K . The interaction of P47 with fibronectin was specific, and the region of fibronectin bound by P47 mapped to the gelatin/collagen binding domain . P47 was purified by affinity chromatography, digested with endoproteinase Lys-C, and the peptide fragments analysed by liquid chromatography/tandem mass spectroscopy . A search of protein databases disclosed that the P47 peptide mass profile matched that predicted for the bbk32 gene product of B . burgdorferi isolate B31 . The bbk32 gene was cloned into Escherichia coli, and the ability of recombinant BBK32 to bind fibronectin and inhibit the attachment of B . burgdorferi was demonstrated . The identification of BBK32 as a receptor for fibronectin binding may enhance our understanding of the pathogenesis and chronic nature of Lyme disease. J Biotechnol, 1999 Jan 8, 67(1), 1 - 11 Patenting in biotechnology: a review of the 20th symposium of ECB8; Szarka E; The patenting of biotechnological inventions is practically in harmony with the general requirements of patent protection . It stands still in the foreground of interests since this is the only technical field where the living material itself may be the subject matter of patents . In consequence ethical problems have arisen first of all in the patenting of human cells and genes in which there is no agreement between R&D firms, patent offices and green movements . This has called for the elaboration of special Directives . On the other hand, patent systems are instrumental in safeguarding biodiversity . This review gives a picture of the patenting situation in biotechnology in the European Patent Office and in Hungary, the host country of the Congress . It also gives practical advice to biotechnological researchers on how to draft the applications and to observe the time limits, as well as on the necessity and possibilities of the deposit of microorganisms. J Clin Microbiol, 1999 Mar, 37(3), 600 - 5 Typing of clinical Mycobacterium avium complex strains cultured during a 2-year period in Denmark by using IS1245; Bauer J et al.; In the present study restriction fragment length polymorphism analyses with the recently described insertion sequence IS1245 as a probe was performed with clinical Mycobacterium avium complex strains cultured in Denmark during a 2-year period . The overall aim of the study was to disclose potential routes of transmission of these microorganisms . As a first step, the genetic diversity among isolates from AIDS patients and non-human immunodeficiency virus (HIV)-infected patients was described . In addition, a number of isolates from nonhuman sources cultured during the same period were analyzed and compared to the human isolates . A total of 203 isolates from AIDS patients (n = 90), non-HIV-infected patients (n = 91), and nonhuman sources (n = 22) were analyzed . The presence of IS1245 was restricted to Mycobacterium avium subsp . avium isolates . The majority of human isolates had large numbers of IS1245 copies, while nonhuman isolates could be divided into a high-copy-number group and a low-copy-number group . Groups of identical strains were found to be geographically widespread, comprising strains from AIDS patients as well as strains from non-HIV-infected patients . Samples of peat (to be used as potting soil) and veterinary samples were found to contain viable M . avium isolates belonging to genotypes also found in humans. Cell Immunol, 1999 Feb 1, 191(2), 117 - 23 Mycobacterium avium complex activates nuclear factor kappaB via induction of inflammatory cytokines; Ghassemi M et al.; A variety of microorganisms has been reported to directly induce NF-kappaB, a critical step in the regulation of genes involved in the cellular immune response . In this study, we demonstrate that proinflammatory cytokines such as tumor necrosis factor alpha (TNFalpha) produced upon activation by the Mycobacterium avium complex (MAC) preceed NF-kappaB activity in U937, a human monocytoid cell line . MAC induction of TNFalpha mRNA expression was detected within 15 min after MAC infection, whereas enhanced NF-kappaB binding activity was not detected until 90 to 120 min postinfection . Supershift analysis revealed increased p50 in the MAC-induced NF-kappaB binding complexes . Consistent with an autocrine mechanism, anti-TNFalpha antibody and dexamethasone, a known cytokine inhibitor, both completely suppressed the effect of MAC on the induction of NF-kappaB . Taken together, these findings suggest that exposure of monocyte cell membranes to MAC induces endogenous TNFalpha, which in turn enhances NF-kappaB binding activity . The rapid induction of TNFalpha may be important in the initial host response to MAC infection . J Immunol, 1999 Feb 1, 162(3), 1611 - 7 CpG oligodeoxynucleotides can circumvent the Th2 polarization of neonatal responses to vaccines but may fail to fully redirect Th2 responses established by neonatal priming; Kovarik J et al.; Neonatal murine responses to a panel of conventional vaccines differ qualitatively from adult responses by a particular polarization toward a Th2 pattern and a frequent limitation of the Th1 and CTL responses required for protection against intracellular microorganisms . In contrast, DNA vaccines induce adult-like Th1/CTL neonatal responses against the same vaccine Ags . In this report, we show that this can be related to their content in unmethylated CpG motifs . Oligodeoxynucleotides (ODN) containing CpG motifs activate neonatal APCs to produce IL-12 in vitro and induce adult-like Th1 responses to tetanus toxoid and measles Ags in vivo, with production of IgG2a-specific Abs and adult-like secretion of IFN-gamma and IL-5 by Ag-specific T cells . However, in spite of their capacity to trigger neonatal B cell proliferation in vitro, CpG-ODN only partially enhanced early life Ab responses . Finally, using Th1-driving CpG-ODN with the boosting dose of a protein vaccine was sufficient to redirect adult but not neonatally primed Th2 responses . These observations could be important for the development of novel vaccines that will have to be effective early in life. J Immunol, 1999 Feb 1, 162(3), 1310 - 7 Comparison of lung dendritic cells and B cells in stimulating naive antigen-specific T cells; Masten BJ et al.; Dendritic cells (DCs) are specialized APCs that are important in priming naive T cells and can be manipulated in vitro and in vivo to enhance immunizations against microorganisms and tumors . A limitation in the development of suitable immunotherapeutic vaccines for the lung is incomplete information on the role of DCs and other potential APCs in the lung in priming naive T cells . In the current study, we analyzed the relative contributions of murine lung DCs and B cells to process and present OVA to naive CD4+ OVA323-339-specific (DO11.10) T cells in vitro . We also examined their expression of MHC class II and accessory molecules before and after maturation in culture . Similar to DCs from other sites, freshly isolated lung DCs can process OVA, spontaneously up-regulate MHC class II and accessory molecules during overnight culture, and stimulate naive T cells in an Ag-specific manner . In contrast, freshly isolated lung B cells were unable to both process and present native OVA . Furthermore, under conditions of limited OVA323-339 peptide exposure, B cells had a significantly diminished capacity to stimulate T cells, and this correlated with a decreased density of both MHC class II and important costimulatory molecules as compared with lung DCs. FASEB J, 1999 Feb, 13(2), 305 - 12 A role for preadipocytes as macrophage-like cells; Cousin B et al.; Several lines of evidence have supported a link betweeen adipose tissue and immunocompetent cells . This link is illustrated in obesity, where excess adiposity and impaired immune function have been described in both humans and genetically obese rodents . In addition, numerous factors involved in inflammatory response are secreted by both preadipocytes and macrophages . Here we show that proliferating preadipocytes in cell lines and primary cultures, develop phagocytic activity toward microorganisms . This is demonstrated by phagocytosis assays and confocal microscopy . This function disappears when preadipocytes stop proliferating and differentiate into adipocytes . After phagocytosis, preadipocytes show microbicide activity via an oxygen-dependent mechanism . In addition, preadipocytes as well as adipocytes are stained with MOMA-2, a marker of monocyte-macrophage lineage, but are negative for specific mature macrophage markers (F4/80 and Mac-1) . These results suggest that preadipocytes could function as macrophage-like cells and raise the possibility of a potential direct involvement of adipose tissue in inflammatory processes. Biosci Biotechnol Biochem, 1998 Nov, 62(11), 2257 - 9 Evidence that a beta-1,4-endoglucanase secreted by Acetobacter xylinum plays an essential role for the formation of cellulose fiber; Koo HM et al.; Cellulose-producing Acetobacter xylinum has been known to secrete a cellulose-hydrolyzing beta-1,4-endoglucanase (CMCax) . When antibodies to recombinant CMCax were added to the culture medium, the formation of cellulose fiber was severely inhibited . Western blot analysis using the antibody showed that this enzyme was secreted into the medium even by a cellulose non-producing strain (Cel-) . These results indicate that beta-1,4-endoglucanase in the medium plays a critical role in the formation of cellulose fiber by the microorganism. Hepatogastroenterology, 1998 Nov-Dec, 45(24), 2270 - 4 Effect of tuftsin on human Kupffer cell; Kubo S et al.; BACKGROUND/AIMS: Kupffer cells are the most important category of reticuloendothelial cells which are critical for host defense in the liver . We investigated the effects of tuftsin (Thr-Lys-Pro-Arg) on human Kupffer cells . METHODOLOGY: Human Kupffer cells were obtained from the livers of patients with colon cancer . Phagocytosis assay was done by microscopic counting of the number of Kupffer cells that engulfed fluorescent particle(s), and the number of the particles engulfed per Kupffer cell when Kupffer cells were incubated with and without tuftsin . Effect of tuftsin on the release of tumor necrosis factor from Kupffer cells was also studied . RESULTS: Phagocytosis was enhanced significantly by tuftsin . The greatest effect on percentage of phagocytic cells was observed at 1.0 microg/ml of tuftsin . The mean number of particles engulfed per Kupffer cell was also increased with tuftsin 1.0 microg/ml . Tumor necrosis factor release was also significantly increased; the greatest effect was observed at 1.0 microg/ml of tuftsin . CONCLUSIONS: Tuftsin enhances phagocytic activity and tumor necrosis factor release of human Kupffer cells, which are advantageous for host defense against invading microorganisms and tumor cells. Int J Lepr Other Mycobact Dis, 1998 Sep, 66(3), 365 - 73 Mycobacterium lepraemurium, a well-adapted parasite of macrophages: I . Oxygen metabolites; Rojas-Espinosa O et al.; We measured the release of reactive oxygen intermediaries {ROI (hydrogen peroxide and superoxide anion)} by murine peritoneal macrophages challenged in vitro with Mycobacterium lepraemurium (MLM), complement-opsonized yeast, M . bovis BCG, M . phlei, or phorbol myristate acetate (PMA) . We found that except for MLM, all of the other materials provoked the release of significant amounts of hydrogen peroxide and superoxide . MLM entered the macrophages without triggering their oxidative metabolism . Pre-infection of macrophages with MLM did not alter these cells' capacity to release the normal amounts of ROI in response to other microorganisms or PMA . Killing of MLM did not revert the macrophages' failure to release ROI upon ingestion of the microorganism, nor were macrophages able to produce these toxic metabolites when pre-incubated in the presence of murine gamma interferon (IFN-gamma) . MLM has several attributes that allow it to survive within macrophages: a) it is a nontoxigenic microorganism (it does not harm its host), b) it resists the harsh conditions of the intraphagolysosomal milieu (a property perhaps dependent on its thick lipidic envelope), and c) it penetrates the macrophages without triggering their oxidative response (thus avoiding the generation of the toxic intermediaries of oxygen) . For these attributes (and others discussed in this paper), we recognize MLM as a highly evolved, well-adapted parasite of macrophages . In addition, the results of the present study prompted the analysis of the biochemical pathways used by MLM and M . bovis BCG to penetrate into their cellular hosts, a subject now under investigation in our laboratory. Adv Biochem Eng Biotechnol, 1999, 63, 169 - 218 Microbial models for drug metabolism; Azerad R; This review describes microbial transformation studies of drugs, comparing them with the corresponding metabolism in animal systems, and providing technical methods for developing microbial models . Emphasis is laid on the potential for selected microorganisms to mimic all patterns of mammalian biotransformations and to provide preparative methods for structural identification and toxicological and pharmacological studies of drug metabolites. Adv Biochem Eng Biotechnol, 1999, 63, 57 - 72 Stereoinversions using microbial redox-reactions; Carnell AJ; This paper aims to provide a summary of the recent literature on the use of redox enzymes to carry out stereoinversion reactions on chiral secondary alcohols . Emphasis has been placed on biotransformations which result in the deracemization of a racemic substrate to give high value synthetic intermediates in a theoretical 100% yield . Most of the biocatalysts which are competent to carry out such transformations are whole cell systems, which contain the necessary cofactor recycling machinery to facilitate this otherwise entropically disfavoured process . The first section deals with deracemization of compounds such as mandelic acid and pantoyl lactone using two microorganisms which display enantiocomplementary stereospecificity . The deracemization of chiral alcohols such as beta-hydroxyesters, aryl ethanols and terminal 1,2-diols with single microorganisms will then be discussed and the influence of growth and reaction conditions on the selectivity observed will be emphasised . Then the ability of several microorganisms to deracemize by double stereoinversion substrates with two stereocentres such as cyclohexan-1,2-diol, cis and trans indan-1,2-diol and pentan-2,4-diol will be presented and some mechanistic rationale proposed . Lastly enzymes known as epimerases which are important in sugar and deoxysugar biosynthesis will be discussed with reference to some recent work on the mechanism of UDP-glucose epimerase. FEMS Microbiol Lett, 1999 Jan 15, 170(2), 363 - 71 Bioaccumulation of heavy metals by fimbrial designer adhesins; Schembri MA et al.; Naturally occurring adhesins bind to specific molecular targets in a lock-and-key fashion due to the composition of the binding domain of the adhesin . By introduction of random peptide libraries in a suitable surface exposed carrier protein it is possible to create and select designer adhesins with novel binding affinities . Type 1 fimbriae are surface organelles of Escherichia coli which mediate D-mannose sensitive binding to different host surfaces through the FimH adhesin, an integral part of these organelles . We have studied the ability of the FimH adhesin to display random peptide sequences . By serial selection and enrichment procedures specific sequences were identified which conferred the ability on recombinant cells to adhere to various metal oxides (PbO2, CoO, MnO2, Cr2O3) . The properties inherent in these sequences permitted the distinct recognition of metals to varying degrees, indicating that this system allow for the isolation of peptide sequences with a variety of binding avidities . These studies demonstrate the potential and versatility of the FimH display system for presenting random peptide sequences . In addition, the possibility exists for the construction of microorganisms for the bioaccumulation of heavy metals from the environment. Clin Exp Immunol, 1999 Feb, 115(2), 229 - 35 Regulation of CR3 (CD11b/CD18)-dependent natural killer (NK) cell cytotoxicity by tumour target cell MHC class I molecules; Vetvicka V et al.; Phagocyte and NK cell CR3 functions as both an adhesion molecule and an iC3b receptor mediating cytotoxic responses to microorganisms . Cytotoxic activation of iC3b receptor function requires ligation of both a CD11b I-domain site for iC3b and a lectin site located in the C-terminus of CD11b . Because tumours lack the CR3-binding polysaccharides of bacteria and fungi, iC3b-opsonized tumours do not stimulate CR3-dependent cytotoxicity . Previous studies showed that NK cells could be induced to kill iC3b-opsonized tumours with small soluble beta-glucans that bound with high affinity to CR3, bypassing the absence of similar polysaccharides on tumour membranes . Because CR3 signalling requires several tyrosine phosphorylation events, it appeared possible that CR3-dependent killing of autologous tumour cells might be suppressed by NK cell inhibitory receptors for MHC class I (KIR and CD94/NKG2) whose action involves recruitment of SHP-1 and SHP-2 tyrosine phosphatases . In the current study, Epstein-Barr virus (EBV)-transformed B cells were used as targets following opsonization with iC3b . Soluble beta-glucan primed CR3 for killing of iC3b-coated B cells, but autologous class I-bearing targets were 84% more resistant than class I-deficient Daudi cells . Blockade of target cell class I with a MoAb specific for a domain recognized by both KIR and CD94/NKG2 resulted in comparable killing of class I+ B cells . By contrast, another MoAb to class II had no effect on cytotoxicity . These data suggest that NK cell recognition of class I suppresses CR3/tyrosine kinase-dependent cytotoxicity in the same way as it suppresses cytotoxicity mediated by other tyrosine kinase-linked receptors such as FcgammaRIIIA (CD16). Am J Ophthalmol, 1999 Jan, 127(1), 77 - 84 Disinfection of tonometers and contact lenses in the office setting: are current techniques adequate? Smith CA, Pepose JS. PURPOSE: To determine whether routine office techniques used to disinfect tonometer prisms and trial contact lenses are sufficient to prevent transmission of ocular infections . METHOD: We reviewed the current literature on the efficacy of certain disinfection protocols against commonly encountered viral, bacterial, and fungal pathogens as well as Acanthamoeba . RESULTS: Some commonly used disinfecting solutions and techniques may be inadequate for disinfection of viruses such as hepatitis C virus and organisms such as Acanthamoeba . When used in accordance with guidelines published by the United States Centers for Disease Control and Prevention (CDC) and the American Academy of Ophthalmology (AAO), 3% hydrogen peroxide is a very effective disinfectant against a wide variety of microorganisms . Specifically, tonometer prisms disinfected by a 5-minute soak in 3% hydrogen peroxide (or 70% isopropyl alcohol or a 1:10 dilution of sodium hypochlorite) are adequately disinfected against most ocular pathogens, with the exception of Acanthamoeba . Trial contact lenses that are disinfected with a 2-hour soak in 3% hydrogen peroxide are effectively rid of all pathogens of concern . After disinfection, rigid lenses should be stored dry, and soft lenses should be stored in a sterile, preserved solution . Repeat disinfection should be routinely performed at 1-month intervals to prevent regrowth of organisms . CONCLUSION: A safe office environment can be maintained by following current CDC recommendations for disinfection, as well as instituting some additional procedures. Langenbecks Arch Chir Suppl Kongressbd, 1998, 115, 439 - 42 {Pathogenesis of pancreatogenic sepsis}; Runkel N; The intestinal tract is the motor of sepsis in the "gut-MOF hypothesis" . Acute pancreatitis causes an early severe reduction of intestinal microcirculation with consequent production of radicals and cytokines damaging intestinal integrity . The intestinal organ dysfunction syndrome results in a breakdown of barrier function and a loss of propulsive activity . This leads to microbial overgrowth and bacterial translocation . This liberates cytokines and causes secondary pancreatic infection after lymphatic and systemic bacterial dissemination . Infected pancreatic necrosis by enteric microorganisms is the main cause of pancreatic sepsis. Gene, 1999 Jan 21, 226(2), 139 - 46 Multiple p450alk (cytochrome P450 alkane hydroxylase) genes from the halotolerant yeast Debaryomyces hansenii; Yadav JS et al.; The halotolerant alkane-assimilating yeast Debaryomyces hansenii was examined for P450 alkane hydroxylase genes known to be required for alkane assimilation in Candida . Four distinct P450alk gene segments and an allelic segment were isolated using PCR based on degenerate primers derived from the CYP52 family of alkane-inducible P450 genes . A screen of a genomic library (15-20kb inserts) constructed for this study, using a probe based on the PCR-isolated segments, yielded seven clones . This has led to the isolation and sequence of two full-length genes DH-ALK1 and DH-ALK2 . These genes, each with an ORF of 1557 bp (519 aa), contained no apparent introns and showed 64% nucleotide sequence homology (61% based on the deduced amino acid sequences) . The deduced proteins had predicted molecular weights of 59,254Da (DH-ALK1) and 59,614Da (DH-ALK2) and have been designated CYP52A12 and CYP52A13 by the P450 Nomenclature Committee . Phylogenetic analysis based on Neighbor Joining Tree showed that DH-ALK1 and DH-ALK2 constitute new genes located on two distinct branches and are most related to the gene CYP52A3 (60% deduced aa homology) and are least related to the gene CYP52C2 (41% deduced aa homology), both of C . maltosa . The isolated genes will provide tools to better understand the diversity of the P450alk family in eukaryotic microorganisms adapted to varied environmental conditions. Rev Clin Esp, 1998 Dec, 198(12), 805 - 9 {Seroepidemiology of Bartonella henselae infection in a risk group}; Blanco Ramos JR et al.; BACKGROUND: Bartonella henselae infections are closely related to a number of clinical conditions of growing interest in our environment . As cats are the reservoir, and their bites and scratches are the main transmission mechanism, we attempted to study the seroepidemiology of the infection caused by this microorganism in a risk group of patients (cat owners) and a control group (blood donors) . PATIENTS AND METHODS: October 1977 . Measurement of antibody titres to B . henselae at different dilutions (breakpoint > or = 1:64) by indirect immunofluorescence (IFI) in 83 cat owners and 85 blood donors . Also, an epidemiologic investigation was carried out (age, sex, alcohol, intake, common cat exposure, cat bite or cat scratch, number and age of cats, bite or presence of fleas, use of repellents, clinical picture consistent with cat scratch disease {CSD}) . RESULTS: Twenty-four cat owners (28.9%) and five blood donors (5.9%) had titres > or = 1:64 (OR: 6.51; 95% CI: 2.32 to 22.9) . Also, the logistic regression analysis showed an association with daily alcohol intake and the previous antecedent of lymph node enlargement of unknown origin among cat owners . CONCLUSIONS: A high prevalence of antibodies to B . henselae was demonstrate among cat owners. Rev Clin Esp, 1998 Dec, 198(12), 789 - 93 {Preliminary report of an epidemic tularemia outbreak in Valladolid}; Bachiller Luque P et al.; We make here a preliminary report of the first 65 cases of tularemia diagnosed at our hospital pertaining to an epidemic outbreak occurred in the area served by the hospital . Diagnosis was made by the presence of a consistent clinical picture and the following laboratory criteria: antibody titre to . Francisella tularensis (tube agglutination) higher than or equal to 1/160, seroconversion or recovery of the microorganism from biological material . The mean age of our patients was 53.8 +/- 13.5 years and female sex predominated (45 women) . Fever, lymph node enlargement, cutaneous ulcers, asthenia, and weight loss were the most common symptoms . The presentation clinical forms included: ulceroganglionar (69%), typhoidal (14%), ganglionar (12.5%), pneumonic (1.5%), oculoganglionar (1.5%) and atypical (1.5%) . The analytical data did not show significant changes with the exception of erythrocyte sedimentation rate . Streptomycin was the antibiotic of choice, followed by ciprofloxacin and gentamicin . The failure rate was 22%, and ciprofloxacin was used for retreatment in twelve occasions and ofloxacin in 1 occasion with a good response . Tularemia is an infective disease which can become endemic in Spain and that should be considered by clinicians, particularly when unilateral enlarged lymph nodes, prolonged febrile syndromes, pharyngitis with negative culture, poor response to beta-lactams and atypical pneumonia are present. Baillieres Clin Rheumatol, 1998 Nov, 12(4), 649 - 64 The spectrum of skin, mucosa and other extra-articular manifestations; Angulo J et al.; The seronegative spondyloarthropathies appear to be the genetically predisposed host's clinical expression to acute, subacute or chronic reaction to the invasion by environmental microorganisms . In the ensuing days or weeks, depending on the infectious load, clinical manifestations may occur ranging from constitutional complaints such as fever, to a variety of symptoms and/or signs related to the portal of entry-intestinal, genitourinary or respiratory . Within weeks or months, the initial or other target organs, such as the mucocutaneous, ocular and cardiovascular systems, may develop an acute reaction of greater or lesser specificity regarding the triggering agent (oral ulcers, circinate balanitis, erythema nodosum, acute anterior uveitis, pericarditis, heart blocks) . Lastly, many years later, a minority of patients, probably those with a large genetic component, exhibit a spectrum of clinical manifestations related to those organs, with a chronic or recurrent course . Acute clinical manifestations--reactive arthritis--are prominent in the initial phase of the clinical spectrum, while chronic manifestations--ankylosing spondylitis--are seen at the other end of the spectrum. Ann N Y Acad Sci, 1998 Nov 17, 859, 65 - 74 Interaction of microorganisms, epithelium, and lymphoid cells of the mucosa-associated lymphoid tissue; Hamzaoui N et al.; Differentiation of specific epithelial cell lineages during development, as well as epithelial plasticity in response to heterologous cell-to-cell cross talk during adult life, accounts for the large variety of functions which are performed by the mucosal surfaces found in the human body . Among its functions, the digestive mucosa is able to sample antigens and microorganisms through M cells of Peyer's patches' follicle-associated epithelium, in order to trigger the development of either tolerance or immune responses . At least in the gut, M-cell formation is immunoregulated . Close contact between immune cells and intestinal epithelium modifies the permeability of the epithelial barrier by inducing the conversion of enterocytes into M cells, offering at the same time an opportunistic way of invasion for pathogens . These lympho-epithelial interactions triggering M-cell formation have now been modeled in culture. Ann N Y Acad Sci, 1998 Nov 17, 859, 37 - 45 Interactions between epithelial cells and immune cells in the intestine; Strober W; Not merely a passive barrier as previously assumed, epithelial cells act as sensitive indicators of infection that initiate defense responses . Noninvasive as well as invasive organisms have been demonstrated to elicit production of chemoattractants . That noninvasive organisms have this capacity suggests that receptor-mediated signaling pathways may be involved . Epithelial cells have been found to release both chemokines and cytokines in a precise mixture that varies according to the origin or maturity of the cell . Different microorganisms also elicit different response patterns . The information presented provides a basis for a new view of epithelial cell function in relation to host defense. Gastroenterol Hepatol, 1998 Dec, 21(10), 479 - 82 {Incidence and clinical features of splenic abscesses, with special reference to tuberculous etiology in a general hospital}; Arruabarrena I et al.; BACKGROUND: The aim of this revision is to know the incidence of splenic abscess (SA) in our hospital, its etiology, with special reference to tuberculosis, and clinical characteristics . PATIENTS AND METHODS: Abdominal CT-scan performed during the period 1987-1997, with the diagnosis of splenic abscess were reviewed . Etiologic diagnosis standed on blood or sputum cultures, PAAF and/or histologic study of lymph nodes . RESULTS: Seventeen cases of SA were obtained, 12 males and 5 females . Limits of age: 13 and 77 years . The causal microorganisms were: M . tuberculosis (7), Mycobacterium aviumintracellulare (1), S . aureus (2), S . anginosus (1), S . milleri (1), E . coli (1), C . albicans (1), T . biguelle (1) and polymicrobian flora (1) . One case was of unknown etiology . Underlying illnesses were: AIDS (7), malignant neoplasms (3), diabetes (2), endocarditis (2), Sjogren syndrome (1) and complications of abdominal surgery (2) . Clinical presentation in nontuberculous splenic abscess was fever and upper-left abdominal pain . Predominant symptoms in tuberculous splenic abscess were fever and weight loss . Blood cultures were positive in 80% of non tuberculous splenic abscess . Specific treatment for tuberculosis improved all patients with tuberculous splenic abscess, without needing surgery or corticosteroids . CONCLUSIONS: From the total of splenic abscess, 41.1% were tuberculous, six with AIDS and one with Sjogren syndrome . Diabetes and malignant neoplasms were the commonest underlying illnesses in the non-tuberculous . In these, clinical presentation consisted in fever and upper-left abdominal pain . In patients with tuberculous splenic abscess, the main complaint was weight loss . A prompt treatment is generally succesful. Insect Mol Biol, 1999 Feb, 8(1), 125 - 32 Tissue tropism, transmission and expression of foreign genes in vivo in midgut symbionts of tsetse flies; Cheng Q et al.; Tsetse flies (Diptera: Glossinidae) harbour three different symbiotic organisms in addition to the pathogenic African trypanosomes they transmit . The two gut-associated symbionts (primary, P; secondary, S) are enteric and are nutritionally required, whereas the third microorganism Wolbachia (family Rickettsiaceae) affects the reproductive biology of the insects it infects . The bacteriome-associated P-symbiont (Wigglesworthia glossinidia) displays a concordant phylogeny with its host tsetse species, whereas midgut S-symbionts characterized from distant tsetse have identical 16S rDNA sequences and therefore may either represent recent independent acquisitions or horizontal transfer between species . The S-symbionts have been cultured in vitro and a genetic transformation system has been developed . Here we report on their density and tissue tropism in different species (G . m . morsitans, G . p . palpalis, G . austeni and G . brevipalpis) and on their maternal route of transmission to tsetse progeny . Using a bacterium-specific PCR-assay, the S-symbionts were found primarily in the midgut, haemolymph, milk gland and in G . palpalis also in salivary glands of teneral flies . In older flies these infections were found to spread to other tissues including muscle, testes and fat body . The S-symbionts were transformed to express the marker gene product, Green Fluorescent Protein (GFP) in vitro . When the recombinant symbionts were introduced into the haemoceal of fertile female flies via intrathoracic microinjection, they were detected in the intrauterine progeny, indicating that haemolymph may provide a possible route for their transmission . The implications of these results for symbiont-host interactions and for transgenic strategies in tsetse are discussed. Appl Environ Microbiol, 1999 Feb, 65(2), 422 - 30 Quantifying microbial diversity: morphotypes, 16S rRNA genes, and carotenoids of oxygenic phototrophs in microbial mats; N bel U et al.; We quantified the diversity of oxygenic phototrophic microorganisms present in eight hypersaline microbial mats on the basis of three cultivation-independent approaches . Morphological diversity was studied by microscopy . The diversity of carotenoids was examined by extraction from mat samples and high-pressure liquid chromatography analysis . The diversity of 16S rRNA genes from oxygenic phototrophic microorganisms was investigated by extraction of total DNA from mat samples, amplification of 16S rRNA gene segments from cyanobacteria and plastids of eukaryotic algae by phylum-specific PCR, and sequence-dependent separation of amplification products by denaturing-gradient gel electrophoresis . A numerical approach was introduced to correct for crowding the results of chromatographic and electrophoretic analyses . Diversity estimates typically varied up to twofold among mats . The congruence of richness estimates and Shannon-Weaver indices based on numbers and proportional abundances of unique morphotypes, 16S rRNA genes, and carotenoids unveiled the underlying diversity of oxygenic phototrophic microorganisms in the eight mat communities studied. Appl Environ Microbiol, 1999 Feb, 65(2), 746 - 51 Isolation of a bacterial strain able to degrade branched nonylphenol; Tanghe T et al.; Conventional enrichment of microorganisms on branched nonylphenol (NP) as only carbon and energy source yielded mixed cultures able to grow on the organic compound . However, plating yielded no single colonies capable, alone or in combination with other isolates, of degrading the NP in liquid culture . Therefore, a special approach was used, referred to as "serial dilution-plate resuspension," to reduce culture complexity . In this way, one isolate, TTNP3, tentatively identified as a Sphingomonas sp., was found to be able to grow on NP in liquid culture . Remarkably, this isolate was able to be filtered through a 0.45-micrometer-pore-diameter filter . Moreover, isolate TTNP3 did not form visible colonies on mineral medium with NP, and it formed visible colonies on R2A agar only after a prolonged incubation of 1 week . High-performance liquid chromatography and gas chromatography-mass spectroscopy analysis of the culture media indicated that the strain starts the degradation of NP with a fission of the phenol ring and preferably uses the para isomer of NP and not the ortho isomer . No distinct accumulation of an intermediary product could be observed. Pathologica, 1998 Oct, 90(5), 463 - 6 Inflammatory pseudotumor of the liver . Report of two cases; Yavuz E et al.; Inflammatory pseudotumor (IPT) is a rare lesion of the liver . We report two cases of IPT of the liver . Patients were a 32-year-old man and a 45-year-old female . Preoperative clinical diagnoses were respectively amebic abscess and metastatic tumor . Fine needle aspirations were performed and cytopathologic findings were compatible with an inflammatory process in both cases . Simple excision was performed in the first case and left lobectomy in the second one . Gross examination revealed expansive solid tumoral masses which measured 3 x 2 x 2 cm and 7 x 5 x 5 cm, respectively . Both lesions were yellow and firm . On microscopic examination, lesions were composed of fascicles of plump spindle cells intermingled with a varying number of plasma cells, lymphocytes and histiocytes . The spindle cell fascicles also contained a moderate amount of intercellular collagen . No microorganism was found on PAS, Giemsa, Grocott and Ziehl Nielsen stained sections . Plasma cells were immunoreactive for both kappa and lambda light chains, revealing their polyclonal nature . Since hepatocellular carcinoma is concerned in differential diagnosis, IPT is of particular clinical importance . Despite the limited experience with fine needle aspiration cytology, histopathologic diagnosis of IPT does not pose great difficulty. J Endod, 1998 Sep, 24(9), 604 - 6 In vitro evaluation of the cytotoxicity and genotoxicity of root canal medicines on human pulp fibroblasts; Chang YC et al.; An intracanal medicine is often required because microorganisms in the dentinal tubes may be difficult to eliminate completely by instrumentation . Phenolic compounds are widely used in dental treatment as sedatives for the dental pulp or as disinfectants for caries and the root canal . In this study, propidium iodide fluorescence and DNA precipitation assay were used to investigated the cytotoxic and genotoxic effects of camphorated phenol and camphorated parachlorophenol on cultured human pulp fibroblasts in vitro . Both medicines reduced the content of double-stranded polynucleic acid of fibroblasts over a 24-h culture period in a concentration-dependent manner . Camphorated parachlorophenol was more cytotoxic than camphorated phenol . But, both medicines did not cause genotoxicity on pulp cells . The advantage of these experimental methods are simplicity and rapidity . Furthermore, this experimental system may be useful for preliminary cytotoxicity and genotoxicity screening of various dental medicines in vitro. Rev Hosp Clin Fac Med Sao Paulo, 1998 Jul-Aug, 53(4), 202 - 4 {Reiter's syndrome and human immunodeficiency virus infection}; de Mello e Silva AC et al.; The increase of rheumatic disorders in HIV infected has been known for the last decade with special emphasis on Reiter's Syndrome . In spite of common sources of HIV and other microorganisms being important in the pathogenesis of Reiter's Syndrome, severe immunodeficiency may be important in the development of rheumatic diseases, mainly because of the presence of arthritogenic microorganisms . The decrease of TCD4 lymphocytes, the increase of TCD8 lymphocytes and the modification of immune regulation may be important in the pathogenesis of Reiter's Syndrome in patients infected by HIV . There is no clinical response to nonhormonal anti-inflammatory drugs in the treatment of this entity. J Prosthet Dent, 1999 Feb, 81(2), 202 - 6 Disinfection of denture base acrylic resin; Lin JJ et al.; STATEMENT OF PROBLEM: During repair or adjustments of acrylic resin removable complete and partial dentures, particles of the acrylic resin from the interior of the prosthesis may expose dental personnel to microbial health hazards if the prosthesis has not been thoroughly disinfected . PURPOSE: This study investigates the efficacy of a commercially prepared microbial disinfectant (Alcide) on the external and internal surfaces of acrylic resins . MATERIAL AND METHODS: Four groups of acrylic resin were incubated in an experimental model to simulate the oral environment over time . Specimens were treated in 2 groups, disinfected and not disinfected, and then further grouped by breaking and not breaking . Analysis was performed with microbial colony counts, SEM, and statistical analyses . RESULTS: Viable microorganisms still remain on the internal and external surfaces of treated resins . CONCLUSION: Chlorine dioxide reduces, but does not eliminate, viable microorganisms on these dental prostheses. Nurs Clin North Am, 1999 Mar, 34(1), 95 - 112 State of the art: antiretroviral and prophylactic treatments in HIV/AIDS; Porche DJ; HIV results in a continual and progressive pathological attack on the human immune system . This attack on the immune system can be altered with combination antiretroviral agents that interrupt the HIV life-cycle . There are three classes of approved antiretroviral agents: nucleoside reverse transcriptase inhibitors, nonnucleoside reverse transcriptase inhibitors, and protease inhibitors . The progressive deterioration of the patient's immune system by HIV infection increases the likelihood of developing opportunistic infections . Primary measures that can be taken to prevent the development of opportunistic infections are preventing exposure to the microorganisms in the environment and maintaining a competent immune system using antiretroviral therapy through prophylactic agents specific to opportunistic infections. Langenbecks Arch Surg, 1998 Dec, 383(6), 437 - 41 A new method to prevent wound infection: a controlled clinical trial in patients with combined liver and bile duct resection; Hiramatsu K et al.; INTRODUCTION: Despite advances in antibiotic prophylaxis, postoperative wound infection remains a major source of morbidity after digestive surgery . Its prevention is a challenging problem, especially in high-risk patients . The authors introduced a new method to prevent surgical wound infections and evaluated its efficacy in a prospective, randomized trial in markedly high-risk patients . METHODS: Patients with biliary tract carcinoma who were scheduled to undergo combined liver and extrahepatic bile duct resection with biliary reconstruction were randomly assigned to one of two groups, well matched in terms of clinical characteristics at baseline . In one group the new treatment was employed (sealed group, n=31), and in the other the wound was treated in the usual fashion (open group, n=28) . In the sealed group, povidone-iodine gel was administered to the subcutaneous tissue, and the skin and peritoneum were approximated with a continuous suture . Wound infection was registered up to 30 days after surgery . RESULTS: Wound infection occurred in 18 patients: 5 (16%) patients in the sealed group and 13 (46%) in the open group (P<0.05) . All 18 underwent preoperative percutaneous transhepatic biliary drainage and had positive bile culture findings . In 13 of these 18 patients (72%) the microorganisms isolated from the infected wound were identical to those in the bile . CONCLUSIONS: Our results confirm the close association between infected bile and wound infection in hepatobiliary surgery . Our new method, "direct wound sealing," is simple, easy to perform, virtually cost-free, and has the potential to prevent wound infections even in markedly high-risk patients. J Biol Chem, 1999 Feb 5, 274(6), 3580 - 9 Molecular determinants of oligomer formation and complement fixation in mannose-binding proteins; Wallis R et al.; Rat serum mannose-binding protein (MBP-A) functions as part of the innate immune system by targetting complement toward potentially pathogenic microorganisms . In order to examine the molecular basis for complement activation, rat MBP-A has been overproduced in Chinese hamster ovary cells . Recombinant protein is post-translationally modified in the same way as the native lectin . Hydrodynamic studies indicate that MBP-A consists predominantly of covalent oligomers containing one to four copies of a subunit that comprises a trimer of polypeptides . These oligomers are non-interconverting and do not assemble into higher order structures at concentrations in excess of those normally found in serum . Disulfide bonds formed between cysteine residues at the N-terminal end of the collagen-like domain link polypeptides to form covalent oligomers . Analysis of wild-type MBP-A and MBP-A containing the substitution Cys6 --> Ser suggests that polypeptides within each trimeric structural unit are mostly linked by disulfide bonds between cysteine residues at positions 13 and 18 arranged in an asymmetrical configuration . Disulfide bonds involving Cys6 connect polypeptides within separate trimers . Analysis of chimeras between MBP-A and rat liver MBP (MBP-C) indicates that residues within the N-terminal region of the collagenous domain and the cysteine-rich domain of MBP-A enable assembly of trimers into higher order oligomers . The activity of MBP-A in a hemolytic complement fixation assay using mannan-coated sheep erythrocytes was approximately 20-fold greater than the activity of MBP-C . Analysis of the MBP chimeras and isolated oligomers of MBP-A reveals that the larger oligomers are more efficient at complement activation . These data indicate that the overall complement fixing activity of MBP-A is a function of the individual molecular activities of oligomers and their relative abundance within the serum. FEMS Microbiol Lett, 1999 Jan 1, 170(1), 271 - 9 Changes in the microbial community in Japan Trench sediment from a depth of 6292 m during cultivation without decompression; Yanagibayashi M et al.; A sample of deep-sea sediment was obtained from the Japan Trench at a depth of 6292 m using a pressure-retaining sediment sampler . Microorganisms in the sediment sample were cultivated in marine broth 2216 at ambient pressure (65 MPa) without decompression, and at atmospheric pressure (0.1 MPa) as a control experiment . 16S ribosomal RNA genes (rDNA) were amplified by PCR from DNA extracted from the original sediment sample and the mixed cultures, and the nucleotide sequences were determined . The results of phylogenetic analysis based on 16S rDNA sequences indicated that microbial diversity in the original sediment samples showed a wide distribution of types in the domain Bacteria . Furthermore, in the mixed cultures incubated at 65 MPa without decompression, bacterial strains belonging to the Shewanella barophiles branch and the genus Moritella existed together at the beginning of cultivation, and Moritella strains became dominant towards the end of the cultivation period . Finally, in the mixed cultures incubated at atmospheric pressure, strains belonging to the genus Pseudomonas were dominant at all times . Analysis of fatty acids extracted from the cultures supported the phylogenetic results. J Antibiot (Tokyo), 1998 Nov, 51(11), 973 - 82 Bioprobes for investigating mammalian cell cycle control; Osada H; Bioprobes are low molecular weight compounds which are useful for investigating mammalian cell functions . The use of bioprobes has substantially assisted the investigation of complex biochemical processes of the mammalian cell cycle . In this review, cell cycle inhibitors mainly isolated from the microorganism are described and their possibility as an antitumor agents is considered . Most cancer cells have some abnormality in the control mechanism of cell cycle progression . Cyclin-dependent kinases (Cdk), which are activated by the binding with the cyclin and simultaneously by the phosphorylation/dephosphorylation of itself, play important roles as engines in the cell cycle . Tubulins are considered to be one of the most important proteins of the cell division machinery . Therefore, Cdk inhibitors and tubulin binders are possible anticancer drugs . Since the function of proteins controlling the cell cycle is also regulated by phosphorylation and dephosphorylation, inhibitors of protein kinases and phosphatases are considered as possible an antitumor agents . We expect that some bioprobes will be developed for clinical use. Pharmacotherapy, 1999 Jan, 19(1), 70 - 5 Cyclospora: review of an emerging parasite; Brown GH et al.; Cyclospora is a parasite traditionally associated with diarrhea in travelers to endemic countries . Recently, several cases of cyclosporiasis were reported in nontravelers in the United States and Canada, implicating various fruits and vegetables as vehicles of infection . The life cycle of cyclospora is not fully known, but is believed to involve both asexual and sexual stages of proliferation . Food- and water-borne transmission of infection have been implicated . Patients infected with Cyclospora cayetanensis have protracted watery diarrhea . Various generalized symptoms are also present, making cyclosporiasis indistinguishable from infectious diarrhea caused by other microorganisms . Diagnosis depends on identifying the organism by microbiologic examination of stool samples . Treatment consists of supportive care, maintenance of fluid and electrolyte status, symptomatic relief, and antibiotic therapy . Trimethoprim-sulfamethoxazole is the only antibiotic available that is effective in eradicating the organism. Infect Immun, 1999 Feb, 67(2), 805 - 9 Intranasal immunization confers protection against murine Pneumocystis carinii lung infection; Pascale JM et al.; To evaluate the feasibility of mucosal immunization against Pneumocystis carinii (Pc) experimental infection, female BALB/c mice were intranasally immunized three times with soluble Pc antigens plus cholera toxin fraction B (Pc-CTB); control groups received either Pc antigen, CTB, or phosphate-buffered saline (PBS) alone . Two weeks after the last immunization, five animals from each group were sacrificed, and cellular and humoral immune responses were evaluated . The remaining five mice were CD4 depleted using a monoclonal antibody against mouse CD4 and inoculated with viable Pc . Significantly higher specific lymphoproliferative responses from tracheobronchial lymph node cells, immunoglobulin M (IgM) and IgG antibody levels in serum, and bronchoalveolar lavage (BAL)-derived IgA antibody concentrations were observed in the Pc-CTB group of mice relative to control groups (P < 0.01) . Five weeks after challenge, no Pc organisms were observed in the lung smears of the Pc-CTB group, while the animals receiving antigen, adjuvant, or PBS had progressively higher numbers of Pc microorganisms . By Western blot analysis, a strongly reactive 55- to 60-kDa antigen was recognized by BAL IgA and by serum IgG . In summary, mucosal immunization elicited specific cellular and humoral immune responses and protected against Pc lung infection after immunosuppression. Infect Immun, 1999 Feb, 67(2), 469 - 77 The mannose receptor mediates uptake of pathogenic and nonpathogenic mycobacteria and bypasses bactericidal responses in human macrophages; Astarie-Dequeker C et al.; The mannose receptor (MR) is involved in the phagocytosis of pathogenic microorganisms . Here we investigated its role in the bactericidal functions of human monocyte-derived macrophages (MDMs), using (i) trimannoside-bovine serum albumin (BSA)-coated latex beads and zymosan as particulate ligands of the MR, and (ii) mannan and mannose-BSA as soluble ligands . We show that phagocytosis of mannosylated latex beads did not elicit the production of O2- . Zymosan, which is composed of alpha-mannan and beta-glucan, was internalized by the MR and a beta-glucan receptor, but the production of O2- was triggered only by phagocytosis through the beta-glucan receptor . Activation and translocation of Hck, a Src family tyrosine kinase located on lysosomes, has previously been used as a marker of fusion between lysosomes and phagosomes in human neutrophils . In MDMs, Hck was activated and recruited to phagosomes containing zymosan later than LAMP-1 and CD63 . Phagosomes containing mannosylated latex beads fused with LAMP-1 and CD63 vesicles but not with the Hck compartment, and the kinase was not activated . We also demonstrate that the MR was unable to distinguish between nonpathogenic and pathogenic mycobacteria, as they were internalized at similar rates by this receptor, indicating that this route of entry cannot be considered as a differential determinant of the intracellular fate of mycobacteria . In conclusion, MR-dependent phagocytosis is coupled neither to the activation of NADPH oxidase nor to the maturation of phagosomes until fusion with the Hck compartment and therefore constitutes a safe portal of entry for microorganisms. Immunol Rev, 1998 Dec, 166, 377 - 84 Ichthyophthirius multifiliis: a model of cutaneous infection and immunity in fishes; Dickerson H et al.; The parasitic ciliate Ichthyophthirius multifiliis offers a useful system for the study of cutaneous immunity against an infectious microorganism . Naive fish usually die following infection, but animals surviving sublethal parasite exposure become resistant to subsequent challenge . This resistance correlates with the presence of humoral antibodies in the sera and cutaneous mucus of immune fish . A mechanism of immunity has recently been elucidated that involves antibody binding to surface proteins (referred to as immobilization antigens or i-antigens) located on the parasite cell and ciliary membranes . Antibody-mediated cross-linking of i-antigens triggers a response by the parasite resulting in its exit from the host . These effects can be observed directly on the surface of live fish . In addition to allowing the observation of effector responses in vivo, Ichthyophthirius also provides a means to study the ontogeny of the mucosal immune response . The sites of antigen capture and presentation, and the sites of antibody production, are unknown with regard to cutaneous immunity . Because the external epithelial surfaces of fish are often the points of pathogen entry, a basic understanding of the inductive immune mechanisms and immune cell interactions in the skin and gills is extremely important with regard to vaccine development . The development of Ichthyophthirius as an experimental system and how it might be used to address these issues are discussed in this review. J Egypt Soc Parasitol, 1998 Dec, 28(3), 729 - 38 Detection of Microsporidia by different staining techniques; Awadalla HN et al.; Previous detection of Microsporidia relied mainly on electron microscopy and histopathology . Recently, non invasive methods were able to recognize this microorganism . In the present study, different stains were used as a means of diagnosing spores of Microsporidia in stool samples of immunosuppressed patients . The original modified trichrome stain (MTS) was used as a standard screening technique for all stool samples . Positive samples for Microsporidia were then stained with the trichrome blue stain, Didier's trichrome blue stain, acid-fast trichrome stain (AFT), modified Ziehl-Neelsen stain, giemsa stain and calcofluor white M2R stain . Both calcofluor and the AFT stains were most efficient . They could simultaneously detect coccidial oocysts and microsporidial spores . This is beneficial and time-saving in the diagnosis of stool samples of immunosuppressed patients, which usually contain more than one opportunistic protozoon . Both stains are easy to perform and require the least amount of staining and examination. Free Radic Biol Med, 1999 Feb, 26(3-4), 482 - 90 Phagocytic killing of microorganisms by radical processes: consequences of the reaction of hydroxyl radicals with chloride yielding chlorine atoms; Saran M et al.; Chloride anions and hydrogen peroxide serve as substrates for myeloperoxidase (MPO) in order to produce hypochlorous acid (HOCl) as one of the major killing agents of phagocytic leukocytes . Apart from this role of being a substrate for the MPO-reaction the chloride anion has been considered as unreactive and has not been implicated in radical reactions which contribute to the killing process . From the inherent reactivities of the pertinent radicals (as determined by pulse radiolysis experiments), the great abundance of chloride, and the most probable distribution of reactants within the phagosome, we deduce estimates for the average life-time and free diffusion path-length in this milieu and arrive at a model according to which chloride ions enter into radical chains and influence the killing of ingested bacteria to an extraordinarily high extent . We propose that hydroxyl radicals--despite some controversial arguments in the literature--may still be considered as important contributors to cell killing especially since we show that their reactions are made more effective by producing chlorine radicals in a cyclic process . We furthermore present arguments how the phagocyte may protect itself from harmful actions of HOCl and H2O2 after the superoxide-generating activity of NADPH oxidase is turned off. Curr Opin Rheumatol, 1999 Jan, 11(1), 24 - 33 The role of superantigens in vasculitis; Tervaert JW et al.; Multiple risk factors are involved in susceptibility to vasculitis . Inherited determinants may increase the risk but are insufficient to induce the disease . Environmental factors, such as infections, are important modulators and probably trigger the disease in most cases . One of the possible triggers may be a bacterial superantigen (SAg) . SAgs may activate autoreactive T cells that mediate autoimmune vessel wall destruction . Furthermore, SAgs may activate autoreactive B cells to produce autoantibodies that are involved in the pathophysiology of vasculitis, such as antineutrophil cytoplasmic autoantibodies or anti-endothelial cell antibodies . In patients with Kawasaki disease, Wegener's granulomatosis, and infection-related forms of vasculitis, SAg-producing microorganisms have regularly been found . Activation of circulating T cells and skewing of the T-cell repertoire have been reported in most forms of vasculitis . In the past year, for the first time, patients were described in which T-cell receptor V beta expansions were documented simultaneously with the typing of the microbial SAgs, providing evidence that the observed changes in the T-cell repertoire could be caused by these bacterial SAgs . In the future, elucidation of the immunologic mechanisms by which SAgs may play a role in the pathophysiology of vasculitis will provide more effective methods for the treatment of vasculitis. Biochimie, 1998 Nov, 80(11), 911 - 21 Genomic organization analysis of acidophilic chemolithotrophic bacteria using pulsed field gel electrophoretic techniques; Amils R et al.; The genomic organization of acidophilic chemolithotrophic bacteria belonging to the genus Thiobacillus, Thiomonas and Leptospirillum was studied using pulsed field gel electrophoresis techniques (PFGE) . The electrophoretic analysis of intact DNA prepared from different strains showed that all have a circular chromosome, with sizes ranging from 1.9 Mb for Leptospirillum ferrooxidans ATCC 49879, the smallest genome for an acidophilic strict chemolithoautotrophic microorganism, to 3.8 Mb for Thiomonas cuprina DSM 5495, the largest in this study . The number of extrachromosomal elements present varied from none, as observed in several isolates of Leptospirillum ferrooxidan, to five in Thiobacillus thiooxidans ATCC 8085 . The mixotroph Thiomonas cuprina DSM 5495 was found to have a linear 50 kb megaplasmid which was inducible when the bacteria was grown in chemolithotrophic conditions . Low-frequency restriction fragment analysis (LFRFA) of different acidophilic chemolithotrophs and related species was carried out by PFGE to determine macrorestriction patterns for rare cutters (SpeI, XbaI, SwaI, PmeI), which were then used for taxonomic identification (karyotyping), genome size determination, and generation of physical and genetic maps. Eur J Med Res, 1999 Jan 26, 4(1), 1 - 7 Ankylosing spondylitis and genitourinary infection; Lange U et al.; One hundred and thirty-four male and 32 female patients with ankylosing spondylitis and 33 women with pure ileitis terminalis Crohn were examined . The study protocol included a medical-rheumatological examination and thorough investigation for genitourinary infection . Urethroadnexitis was found in 37/134 male patients (2 patients suffered from balanitis, 17 patients from urethritis, 18 patients from prostatitis, and 2 patients from epididymitis), 15/32 female patients (11 of them had urethritis and in 4 cases urethritis associated with vaginitis) and 5/33 women with ileitis terminalis (every case with urethritis) . The microorganism isolated most frequently from patients with genitourinary infection was Chlamydia trachomatis . The majority of patients with genitourinary infection were HLA-B27 positive . Nevertheless, the following conclusions can be reached: (1) evidence of Chlamydia trachomatis infection is frequent in male and female patients with ankylosing spondylitis, (2) patients with genitourinary infection tend to have HLA-B27, and (3) furthermore, presence of genitourinary infection was not significantly associated with chronic illness. Ann Allergy Asthma Immunol, 1998 Dec, 81(6), 585 - 92 Serum IgG antibodies against Wallemia sebi and Fusarium species in Finnish farmers; Lappalainen S et al.; BACKGROUND: Wallemia sebi and Fusarium species are common fungi in agricultural environments . Because Fusarium species are difficult to culture, and W . sebi has only recently been found in an agricultural environment with the use of a new culture medium, immunochemical methods may be important for evaluating exposure to these fungi . OBJECTIVE: Immunoglobulin (Ig) G antibodies against W . sebi and two Fusarium species were measured in the sera of Finnish farmers to estimate exposure to fungi . METHODS: Serum-specific IgG antibody levels against the fungi were determined in farmers with farmer's lung disease and asthma, as well as in asymptomatic farmers with low and high IgG antibody levels against other agricultural microorganisms, and in control persons (printing and office workers) . Both enzyme-linked immunosorbent assay (ELISA) and enzyme-linked immunofiltration assay (ELIFA) techniques were used . RESULTS: Significantly higher IgG antibody levels against the fungi studied were observed in farmer's lung patients and asymptomatic farmers with high IgG levels against other agricultural fungi, than in the control group . On the other hand, the development of IgG antibodies to the fungi in asthmatic farmers remained unclear because of a discrepancy between the ELISA and ELIFA results . Cross-reactivity between Fusarium species was detected . CONCLUSIONS: The results indicate that Finnish farmers are exposed to W . sebi and Fusarium species in their work environment to a greater extent than has earlier been evaluated with air sampling and microbiologic methods. Annu Rev Microbiol, 1998, 52, 779 - 806 Cooperative organization of bacterial colonies: from genotype to morphotype; Ben-Jacob E et al.; In nature, bacteria must often cope with difficult environmental conditions . To do so they have developed sophisticated cooperative behavior and intricate communication pathways . Utilizing these elements, motile microbial colonies frequently develop complex patterns in response to adverse growth conditions on hard surfaces under conditions of energy limitation . We employ the term morphotype to refer to specific properties of colonial development . The morphologies we discuss include a tip-splitting (T) morphotype, chiral (C) morphotype, and vortex (V) morphotype . A generic modeling approach was developed by combining a detailed study of the cellular behavior and dynamics during colonial development and invoking concepts derived from the study of pattern formation in nonliving systems . Analysis of patterning behavior of the models suggests bacterial processes whereby communication leads to self-organization by using cooperative cellular interactions . New features emerging from the model include various models of cell-cell signaling, such as long-range chemorepulsion, short-range chemoattraction, and, in the case of the V morphotype, rotational chemotaxis . In this regard, pattern formation in microorganisms can be viewed as the result of the exchange of information between the micro-level (the individual cells) and the macro-level (the colony). APMIS, 1998 Nov, 106(11), 1041 - 8 Quantification of Bordetella pertussis in clinical samples by colorimetric detection of competitive PCR products; Erlandsson A et al.; Quantification of microorganisms is an important part of the normal diagnostic work of a clinical microbiology laboratory . Traditionally the diagnosis of pertussis is subject to a yes or no approach with no quantitative dimension . This can, however, be of interest as a factor when judging the risk of a patient spreading the bacterium and as a research tool . The aim of the present study was to develop a PCR-based quantitative assay for Bordetella pertussis DNA in clinical nasopharyngeal aspirates by combining a quantitative PCR with a colorimetric detection principle, DIANA (detection of immobilised amplified nucleic acid) . A competitor to the PCR target sequence in IS-481, containing a lac-operator, was constructed and calibrated, and a test protocol prepared . A total of 46 clinical nasopharyngeal aspirates, previously diagnosed using a standard nested PCR assay and quantified by culture, were analysed by the quantitative PCR . The method showed acceptable precision and accuracy considering that it estimates the total number of bacterial genomes while culture detects viable bacteria . Recognised advantages were the simple colorimetric detection, the inborn indication of a working PCR assay, and the possibility of obtaining results even when partial inhibition of the PCR assay was seen . In addition, the quantitative PCR result can be obtained within one day compared to 3-10 days for culture . The present results and the qualities of the quantitative PCR suggest that this assay will be a useful complement in routine diagnostics and in research. Biotechnol Annu Rev, 1998, 4, 75 - 112 Modified hemoglobin blood substitutes: present status and future perspectives; Chang TM; Biotechnological techniques of cross-linking and microencapsulation of hemoglobin result in blood substitutes that can replace red blood cells . Unlike red blood cells they can be sterilized by pasteurization, ultrafiltration and chemical means . This removes microorganisms responsible for AIDS, hepatitis, etc . Since they are free of red blood cell blood group antigens, there is no need for cross-matching or typing . This saves time and facilities and allows on-the-spot transfusion such as the infusion of salt solution . Furthermore, they can be stored for a long time . Hemoglobin for modification can be extracted from human red blood cells . Other sources of hemoglobin include bovine hemoglobin and recombinant human hemoglobin . Clinical trials are ongoing testing the possible uses of cross-linked hemoglobin in cardiac, orthopedic, trauma and other types of surgery . It is also being tested for the replacement of lost blood in severe bleeding due to trauma or other causes . Cross-linked hemoglobins are first generation blood substitutes that only fulfil some of the functions of red blood cells . New generations of more complete red blood cell substitutes are being developed . These include cross-linked hemoglobin-catalase-superoxide dismutase and microencapsulated hemoglobin-enzyme systems. Wien Klin Wochenschr, 1998 Nov 27, 110(22), 796 - 801 Receptor and non-receptor mediated formation of superoxide anion and hydrogen peroxide in neutrophils of intensive care patients; Manhart N et al.; Generation of reactive oxygen intermediates (ROI) has been implicated in tissue damage in a variety of disease states including sepsis and trauma . On the other hand, generation of ROI in polymorphonuclear granulocytes (PMN) presents a crucial element in the defence of the host against invading microorganisms . In the present study we investigated the generation of superoxide anions (O2-) and hydrogen peroxide (H2O2) by neutrophils (PMN)5 of 17 critically ill patients treated at a intensive care unit (ICU) after polytrauma (n = 6), heart operation (n = 6) or during septic shock (n = 5) using flow cytometry . O2- production of PMN from ICU patients was significantly lower (p < 0.01) than that in healthy volunteers (HV) during non-receptor mediated stimulation with phorbol-myristate-acetate (PMA) but higher (p < 0.001) during receptor mediated stimulation with formylmethionine-leucine-phenylalanine (FMLP) . H2O2 generation of PMN from ICU patients was increased after stimulation with FMLP (p < 0.01) and remained unchanged after stimulation with PMA . Patients in septic shock had lower O2(-)-generation of PMN than did injured patients and patients after heart operations . We conclude that receptor mediated formation of O2- and H2O2 is stimulated in ICU patients . However, in patients in septic shock O2(-)-generation decreases, which potentially might contribute to the immunoparalysis present in septic shock. Minerva Stomatol, 1998 Nov, 47(11), 613 - 6 {Benign maxillofacial mucormycosis . Report of a case and review of the literature}; Cangiano G et al.; Mucormycosis is a rare and often lethal infection caused by an opportunistic fungus of the Phycomycetes class . This microorganism is a saprophytic aerobial fungus common in nature and in oral mucosae, nose, paranasal sinus and throat . Mucormycosis has been described in various clinical forms: rhinocerebral, pulmonary, systemic, cerebral and cutaneous, always in immunodepressed patients, with rapid evolution and high rate of mortality . However, rare cases involving maxillofacial area are described in healthy patients with benign evolution and good prognosis as in a case observed at the Department of Maxillofacial Surgery of the University of Naples "Federico II". Biochem Biophys Res Commun, 1998 Dec 18, 253(2), 407 - 14 A region of the cellobiohydrolase I promoter from the filamentous fungus Trichoderma reesei mediates glucose repression in Saccharomyces cerevisiae, dependent on mitochondrial activity; Carraro DM et al.; The upstream activating region that controls cellulose-induced expression of the glucose-repressible cellobiohydrolase I gene (UARcb1) of the filamentous fungus Trichoderma reesei is shown to mediate transcription and glucose repression of a reporter gene in Saccharomyces cerevisiae, a unicellular microorganism that lacks the genes required for the utilization of cellulose . Glucose-controlled transcription mediated by UARcb1 requires the products of the genes SNF1 and SSN6, a protein kinase and a repressor, respectively, that regulate glucose-repressible yeast genes . Previously, it has been shown that mitochondrial function is implicated in cellobiohydrolase I gene expression in T . reesei and this sensitivity to the metabolic state of the mitochondria was shown to be transcriptionally controlled by the 5'-flanking sequence of the cbh1 gene {Abrahao-Neto et al . (1995) Biochemistry 34, 10456-10462} . Remarkably, transcription of the reporter gene controlled by UARcb1 in S . cerevisiae also showed a requirement for active mitochondria, suggesting that a common mechanism involving mitochondrial activity controls glucose-repressible genes in both microorganisms . Anim Reprod Sci, 1998 Dec 31, 54(2), 65 - 73 Sanitary status of oocytes and embryos collected from heifers experimentally exposed to Leptospira borgpetersenii serovar hardjobovis; Bielanski A et al.; In a preliminary trial and three experiments, a total of 30 Holstein heifers were experimentally infected with a culture of Leptospira borgpetersenii serovar hardjobovis via one or more routes (uterine, cervical supraconjunctival, intranasal) and oviductal and uterine fluids recovered post-mortem or in vivo following superovulation with FSH . All routes of administration were effective in establishing Leptospira infection in the reproductive tract and Leptospira were identified in the oviductal and uterine fluids of all 30 heifers by microscopy . The incidence of infection was confirmed by positive identification of serum antibodies by the microscopic agglutination test (MAT) . Twenty-one samples of the embryos (n = 59) recovered were cultured using bacteriological procedures and all tested negative for the infectious microorganism . Using polymerase chain reaction (PCR) assay, however, showed that 29% (7/24) of morula and blastocyst stage embryos, and one out of 29 oocytes tested positive for the presence of leptospiral DNA . A single oocyte or embryo collected from the infected heifers was inoculated intravenously to 26 test heifers . None of the test heifers developed antibody titers to Leptospira . It was concluded that, despite the presence of leptospires in the reproductive tract of donor animals and the association of leptospiral DNA with uterine stage embryos, the transmission of this disease is unlikely to occur by transfer of in vivo produced embryos in the bovine. Drug Dev Ind Pharm, 1998 Jul, 24(7), 589 - 97 Recent approaches in insulin delivery; Trehan A et al.; Insulin remains indispensable in the management of diabetes mellitus since its discovery in 1921 . The foreignness of early available porcine and bovine insulin led to the development of human insulin by transpeptidation and biosynthesis in microorganisms . Needle phobia and stress of multiple daily injections led to the investigation and exploitation of all promising routes, ranging from nasal to rectal, by a wide variety of devices and